Four potent, synthetic inhibitors of matrix metalloproteinases (MMPs) were assessed as inhibitors of tumor growth and spontaneous metastasis to the lung. Mat Ly Lu rat prostate tumor, LOX human melanoma and M27 murine Lewis lung tumor were implanted subcutaneously (s.c.) in mice and allowed to grow for 3–12 days. The lungs of the tumor-bearing mice were then removed and implanted s.c. into untreated mice, and the outgrowth of secondary tumors from the implanted lungs measured. The incidence and rate of outgrowth of secondary tumors increased with the length of primary tumor growth, validating these measurements as indices of spontaneous metastasis to the lung. Compounds were tested by sc. implantation of minipumps which delivered compound throughout the period of primary tumor growth and spontaneous metastasis to the lung at steady-state drug concentrations orders of magnitude greater than the concentrations needed to either inhibit collagenase, gelatinase or stromelysin in vitro. Inhibitor treatment slowed the growth of primary s.c. Mat Ly Lu and LOX tumors by 40–60% but had no significant effect on the growth of primary M27 tumors. Surprisingly, inhibitor treatment had no significant effect on the ability of the lung to generate secondary tumors when reimplanted s.c. in untreated mice. Because of the possible importance of cathepsins B, H and L in tumor growth and metastasis, the irreversible inhibitor E-64 was also infused by s.c. minipump. E-64 had no effect on the growth or spontanous metastasis of Mat Ly Lu or M27 tumors.
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References
Folkman, J and Shing, Y, 1992, Angiogenesis. J Biol Chem, 16, 10931–4.
Liotta, LA, 1986, Tumor invasion and metastasis: role of the extracellular matrix. Cancer Res, 46, 1–7.
Ray, JM and Stetler-Stevenson, WG, 1994, The role of matrix metalloproteinases and their inhibitors in tumour invasion, metastasis and angiogenesis. Eur Respir J, 7, 2062–72.
Sloane, BF, Moin, K, Krepela, E and Rozhin, J, 1990, Cathepsin B and its endogenous inhibitors: their role in tumor malignancy. Cancer Metastasis Rev, 9, 333–52.
Wolf, C, Chenard, M, Durand de, Grossouvre P, Bellocq, J, Chambon, P and Basset, P, 1992, Breast cancer-associated stromelysin-3 gene is expressed in basal cell carcinoma and during cutaneous wound healing. J Invest Dermatol, 99, 870–2.
Pyke, C, Ralfkiaer, E, Huhtala, P, Hurskainen, T, Dano, K and Tryggvason, K, 1992, Localization of messenger RNA for Mr 72,000 and 92,000 type IV collagenases in human skin cancers by in situ hybridzation. Cancer Res, 52, 1336–41.
Polette, M, Clavel, C, Cockett, M, Girod De, Bentzmann S, Murphy, G and Birembaut, P, 1993, Detection and localization of matrix metalloproteinases and their tissue inhibitor in human breast pathology. Invasion Metastasis, 13, 31–7.
Stearns, ME and Wang, M, 1993, Type IV collagenase (Mr 72,000) expression in human prostate: benign and malignant tissue. Cancer Res, 53, 878–83.
Wolf, C, Rouyer, N, Lutz, Y, et al. 1993, Stromelysin 3 belongs to a subgroup of proteinases expressed in breast carcinomal fibroblastic cells and possibly implicated in tumor progression. Proc Natl Acad Sci USA, 90, 1843–7.
Kawame, H, Toshida, K, Ohsaki, A, Kuroi, K, Nishiyama, M and Tage, T, 1993, Stromelysin-3 mRNA expression and malignancy: comparison with clinicopathogical features and type IV collagenase mRNA expression in breast tumors. Anticancer Res, 13, 2319–24.
Garbisa, S, Pozzatti, R, Muschel, RJ, et al. 1987, Secretion of type IV collagenolytic protease and metastatic phenotype: induction of transfection with c-Ha-ras but not c-Ha-ras plus Ad2-Ela. Cancer Res, 47, 1523–8.
Sreenath, T, Matrisian, LM, Stetler-Stevenson, W, Gattoni-Celli, S and Pozzatti, RO, 1992, Expression of matrix metalloproteinase genes in transformed rat cell lines of high and low metastatic potential. Cancer Res, 52, 4942–7.
Bernhard, EJ, Muschel, RJ and Hughes, EN, 1990, Mr 92,000 Gelatinase releases correlates with the metastatic phenotype in transformed rat embryo cells. Cancer Res, 50, 3872–7.
Terranova, VP, Hujanene, ES, Loeb, DM, Martin, GR, Thornburg, L and Glushko V, 1986, Use of a reconstituted basement membrane to measure cell invasiveness and select for highly invasive tumor cells. Proc Natl Acad Sci USA, 83, 465–9.
Nakajima, M, Welch, DR, Belloni, PN and Nicolson, GL, 1987, Degradation of basement membrane type IV collagen and lung subendothelial matrix by rat mammary adenocarcinoma cell clones of differing metastatic potentials. Cancer Res, 47, 4869–76.
Schultz, RM, Silberman, S, Persky, B, Bajowski, AS and Carmichael, DF, 1988, Inhibition by human recombinant tissue inhibitor of metalloproteinases of human amnion invasion and lung colonization by murine B16-F10 melanoma cells. Cancer Res, 48, 5539–45.
Alvarez, OA, Carmichael, DF and DeClerck, YA, 1990, Inhibition of collagenolytic activity and metastasis of tumor cells by a recombinant human tissue inhibitor of metalloproteinases. J Natl Cancer Inst, 82, 589–95.
Khokha, R, 1994, Suppression of the tumorigenic and metastatic abilities of murine B 16-F 10 melanoma cells in vivo by the overexpression of the tissue inhibitor of the metalloproteinases-1. J Natl Cancer Inst, 86, 299–304.
DeClerck, YA, Perez, N, Shimada, H, Boone, TC, Langley, KE and Taylor, SM, 1992, Inhibition of invasion and metastasis in cells transfected with an inhibitor of metalloproteinases. Cancer Res, 52, 701–8.
Reich, R, Thompson, EW, Iwanoto, Y, et al. 1988, Effects of inhibitors of plasminogen activator, serine proteinases, and collagenase IV on the invasion of basement membranes by metastatic cells. Cancer Res, 48, 3307–12.
Naito, K, Kanbayashi, N, Nakajima, S, et al. 1994, Inhibition of growth of human tumor cells in nude mice by a metalloproteinase inhibitor. Int J Cancer, 58, 730–5.
Chirivi, RGS, Garofalo, A, Crimmins, MJ, et al. 1994, Inhibition of the metastatic spread and growth of B16-BL6 murine melanoma by a synthetic matrix metalloproteinase inhibitor. Int J Cancer, 58, 460–4.
Wang, X, Fu, X, Brown, PD, Crimmin, MJ and Hoffman, RM, 1994, Matrix metalloproteinase inhibitor BB-94 (Batimastat) inhibits human colon tumor growth and spread in a patient-like orthotopic model in nude mice. Cancer Res, 54, 4726–8.
Davies, B, Brown, PD, East, N, Crimmin, MJ and Balkwill, FR, 1993, A synthetic matrix metalloproteinase inhibitor decreases tumor burden and prolongs survival of mice bearing human ovarian carcinoma xenografts. Cancer Res, 53, 2087–91.
Bickett, DM, Green, MD, Berman, J, et al. 1993, A high throughput fuorogenic substrate for interstitial collagenase (MMP-1) and gelatinase (MMP-9). Anal Biochem, 212, 58–64.
Weingarten, H, Martin, R and Feder, J, 1985, Synthetic substrates of vertebrate collagenases. Biochemistry, 24, 6730–4.
Mosmann, T, 1983, Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods, 65, 55–63.
Campion C, Dickens JP and Crimmin MJ, 1990, Patent WO 90/05719, British Biotechnology.
Porter JR, Morphy JR, Millican TA and Beeley NRA, 1993, New sulphamide derivatives are metalloproteinase inhibitors for inhibiting tumor metastasis. Patent WO 9324475-A1, Cell Tech Ltd.
Barrett, AJ, Kembhavi, AA, Brown, MA, et al. 1982, l-trans-Epoxysuccinyl-leucylamido(4-guanidino)butane (E-64) and its analogues as inhibitors of cysteine proteinases including cathepsins B, H, and L. Biochem J, 201, 189–98.
Dykes D, Shoemaker R, Harrison S, et al. 1987, Development and therapeutic response of a spontaneous metastasis model of a human melanoma. Proc Am Assoc Cancer Res, 28, Abstract 1709.
Hashida, S, Towatari, T, Kominami, E and Katunuma, N, 1980, Inhibitions by E-64 derivatives of rat liver cathepsin B and cathepsin L in vitro and in vivo. J Biochem, 88, 1805–11.
Miyagi, E, Yasumitsu, H, Hirahara, F, et al. 1995, Marked induction of gelatinases, especially type B, in host fibroblasts by human ovarian cancer cells in athymic mice. Clin Exp Metastasis, 13, 89–96.
Conway, JG, Wakefield, JA, Brown, RH, et al. 1995, Inhibition of cartilage and bone destruction in adjuvant arthritis in the rat by a matrix metalloproteinase inhibitor. J Exp Med, 182, 449–57.
Galardy, RE, Grobelny, D, Foellmer, HG and Fernandez, LA, 1994, Inhibition of angiogenesis by the matrix metalloproteinase inhibitor N-[2R-2-(hydroxamidocarbonymethyl) - 4 - (methylpentanoyl)] - l - tryptophan methylamide. Cancer Res, 54, 4715–18.
Taraboletti, G, Garafalo, A, Belotti, D, et al. 1995, Inhibition of angiogenesis and murine hemagioma growth by batimastat, a synthetic inhibitor of matrix metalloproteinases. J Natl Cancer Inst, 87, 293–8.
Dykes, DJ, Griswold, DP, Jr and Schabel, FM, 1976, Growth support of small B16 melanoma implants with nitrourea-sterilized fractions of the same tumor. Cancer Res, 36, 2031–4.
Hewitt, HB, Blake, E and Porter, EH, 1973, The effect of lethally irradiated cells on the transplantability of murine tumours. Br J Cancer, 28, 123–35.
DeWys, WD, 1972, A quantitative model for the study of the growth and treatment of a tumor and its metastasis with correlation between proliferative state and sensitivity to cyclophosphamide. Cancer Res, 32, 367–73.
Wilcox, D and Mason, R, 1992, Inhibition of cysteine proteinases in lysosomes and whole cells. Biochem J, 285, 495–502.
Ostrowski, LE, Ahsan, A, Suthar, BP, et al. 1986, Selective inhibition of proteolytic enzymes in an in vivo mouse model for experimental mestastasis. Cancer Res, 46, 4121–8.
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Conway, J.G., Trexler, S.J., Wakefield, J.A. et al. Effect of matrix metalloproteinase inhibitors on tumor growth and spontaneous metastasis. Clin Exp Metast 14, 115–124 (1996). https://doi.org/10.1007/BF00121208
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DOI: https://doi.org/10.1007/BF00121208